HGF is a protein that enhances proliferation of liver parenchyma cells, and proteins having different amino acid sequences have been reported, and are known in the names of HGF, TCF, SCF, etc. In the invention, these known proteins having hepatocyte growth activity are collectively called HGF.
HGF is a physiological active peptide showing various pharmacological actions, and its pharmacological actions are reported, for example, in Experimental Medicine (Japan), Vol. 10, No. 3 (extra issue), 330–339 (1992). Owing to its pharmacological actions, HGF is expected to be developed as agent for liver cirrhosis, agent for kidney disease, epithelial cell growth promoter, carcinostatic agent, side effect inhibitor for cancer therapy, agent for lung disorder, agent for gastroduodenal lesion, agent for cerebral and nervous disorder, agent for relieving side effects caused by immunosuppressants, collagen decomposition promoter, agent for cartilage disorder, agent for arterial disease, agent for lung fibroid, agent for liver disease, agent for abnormal blood clotting, agent for hypoproteinemia, wound cure agent, improving agent for nervous disorder, hematopoietic stem cell promoter, hair growth promoter, etc. (Japanese Laid-open Patent No. 4-18028, Japanese Laid-open Patent No. 4-49246, EP No. 492614, Japanese Laid-open Patent No. 6-25010, WO 93/8821, Japanese Laid-open Patent No. 6-172207, Japanese Laid-open Patent No. 7-89869, Japanese Laid-open Patent No. 6-40934, WO 94/2165, Japanese Laid-open Patent No. 6-40935, Japanese Laid-open Patent No. 6-56692, Japanese Laid-open Patent No. 7-41429, WO 93/3061, Japanese Laid-open Patent No. 5-213721, etc.).
Preparations of HGF are disclosed in WO 90/10651 and Japanese Laid-open Patent No. 6-247872. This publication of WO 90/10651 discloses a deletion type HGF (dLeHGF) deleting five residues of amino acid from HGF, and it is named TCFII. This specification shows that HGF is stabilized by albumin, human serum, gelatin, sorbitol, mannitol, xylitol, etc. But, it relates to aqueous solution preparations, and HGF is stabilized in an aqueous solution. The publication of Japanese Laid-open Patent No. 6-247872 unveils a preparation having HGF contained at high concentration by coexistence of basic amino acids and HGF (TCF).
Generally, the protein is not so stable in freezing operation (Protein, Nucleic Acid, Enzyme (Japan), 37(9), 1517, 1992). The stabilizer of protein in an aqueous solution is intended to stabilize by mutual action of water molecule and protein. Therefore, in a lyophilized preparation of protein in the absence of water, the stabilizer of protein for an aqueous solution shows no stabilizing effect in most cases (Protein, Nucleic Acid, Enzyme (Japan), 37(9), 1517, 1992).
On the other hand, nothing has been known about lyophilized HGF preparation, and it could not expected how far the lyophilized HGF preparation would show physical and biological stability.
The aqueous solution preparation of HGF itself is, when stored at low temperature or room temperature for several days, changed in properties, showing aggregation, turbidity and gelation, and forms variants and polymers, and it is low in physical stability and is lowered in biological activity, and hence it is low in stability of biological activity and is not a stable preparation suited long-term storage. It has been a fatal point for development of HGF as medicines or animal drugs in a form of injection preparation. The invention solves the above-mentioned problems. That is, it is an object of the invention to present a stable preparation which can store for a long period as medicines for medical treatment or animal drugs.